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Abstract: FR-OR47

Normothermic Ex Vivo Kidney Perfusion in a Porcine Auto-Transplantation Model Preserves the Expression of Key Mitochondrial Proteins: An Unbiased Proteomics Analysis

Session Information

Category: Transplantation

  • 1901 Transplantation: Basic

Authors

  • McEvoy, Caitriona M., Toronto General Hospital Research Institute, University Health Network, Toronto, Ontario, Canada
  • Clotet Freixas, Sergi, Toronto General Hospital Research Institute, University Health Network, Toronto, Ontario, Canada
  • Tokar, Tomas, Krembil Research Institute, Toronto Western Hospital, University Health Network,, Toronto, Ontario, Canada
  • Pastrello, Chiara, Krembil Research Institute, Toronto Western Hospital, University Health Network,, Toronto, Ontario, Canada
  • Reid, Shelby, Institute of Medical Science, University of Toronto,, Toronto, Ontario, Canada
  • Batruch, Ihor, Department of Laboratory Medicine and Pathobiology, Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, University of Toronto, Toronto, Ontario, Canada
  • RaoPeters, Adrien A E, Department of Physiology and Pharmacology, Schulich School of Medicine & Dentistry, Western University, London, Ontario, Canada
  • Urbanellis, Peter, Toronto General Hospital Research Institute, University Health Network, Toronto, Ontario, Canada
  • Farkona, Sofia, Toronto General Hospital Research Institute, University Health Network, Toronto, Ontario, Canada
  • Van, Julie Anh Dung, Institute of Medical Science, University of Toronto,, Toronto, Ontario, Canada
  • Urquhart, Brad, Department of Physiology and Pharmacology, Schulich School of Medicine & Dentistry, Western University, London, Ontario, Canada
  • John, Rohan, Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, Canada
  • Robinson, Lisa, Division of Nephrology, The Hospital for Sick Children, Toronto, Ontario, Canada
  • Selzner, Markus, Soham and Shaila Ajmera Family Transplant Centre, Toronto General Hospital, University Health Network, Toronto, Ontario, Canada
  • Konvalinka, Ana, Toronto General Hospital Research Institute, University Health Network, Toronto, Ontario, Canada
Background

Normothermic ex-vivo kidney perfusion(NEVKP) results in significantly improved graft function in porcine auto-transplant models of DCD injury compared to static cold storage (SCS); however, the molecular mechanisms underlying these beneficial effects remain unclear.

Methods

We performed an unbiased proteomics analysis of 28 kidney biopsies obtained at 3 timepoints from pig kidneys subjected to 30-minutes of warm ischemia, followed by 8 hours of NEVKP or SCS, and auto-transplantation.

Results

Of 6593 proteins quantified, 70 were differentially expressed between NEVKP and SCS groups (2-way ANOVA, q<0.05). Proteins increased in NEVKP mediated key metabolic processes including fatty acid ß-oxidation, the TCA cycle and oxidative phosphorylation. Comparison of our findings with external datasets of ischemia-reperfusion, and other models of kidney injury confirmed that 47 of our proteins represent a common signature of kidney injury reversed or attenuated by NEVKP. We validated key metabolic proteins (ETFB, CPT2) by immunoblotting. Integrated transcription factor databases identified PPARGC1A, PPARA/G/D and RXRA/B as the upstream regulators of our dataset, and we confirmed their increased expression in NEVKP with RT-PCR.

Conclusion

The proteome-level changes observed in NEVKP mediate critical metabolic pathways that may explain improved graft function with NEVKP compared to SCS. These effects may be coordinated by PPAR-family transcription factors, and may represent novel therapeutic targets in ischemia-reperfusion injury.